Application of long-term cultured interferon-γ enzyme-linked immunospot assay for assessing effector and memory T cell responses in cattle

Mayara F. Maggioli*, Mitchell V. Palmer, H. Martin Vordermeier, Adam O. Whelan, James M. Fosse, Brian J. Nonnecke, W. Ray Waters

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Abstract

Effector and memory T cells are generated through developmental programing of naïve cells following antigen recognition. If the infection is controlled up to 95 % of the T cells generated during the expansion phase are eliminated (i.e., contraction phase) and memory T cells remain, sometimes for a lifetime. In humans, two functionally distinct subsets of memory T cells have been described based on the expression of lymph node homing receptors. Central memory T cells express C-C chemokine receptor 7 and CD45RO and are mainly located in T-cell areas of secondary lymphoid organs. Effector memory T cells express CD45RO, lack CCR7 and display receptors associated with lymphocyte homing to peripheral or inflamed tissues. Effector T cells do not express either CCR7 or CD45RO but upon encounter with antigen produce effector cytokines, such as interferon-γ. Interferon-γ release assays are used for the diagnosis of bovine and human tuberculosis and detect primarily effector and effector memory T cell responses. Central memory T cell responses by CD4+ T cells to vaccination, on the other hand, may be used to predict vaccine efficacy, as demonstrated with simian immunodeficiency virus infection of non-human primates, tuberculosis in mice, and malaria in humans. Several studies with mice and humans as well as unpublished data on cattle, have demonstrated that interferon-γ ELISPOT assays measure central memory T cell responses. With this assay, peripheral blood mononuclear cells are cultured in decreasing concentration of antigen for 10 to 14 days (long-term culture), allowing effector responses to peak and wane; facilitating central memory T cells to differentiate and expand within the culture.

Original languageEnglish
Article numbere52833
Number of pages8
JournalJournal of Visualized Experiments
Volume2015
Issue number101
DOIs
Publication statusPublished - 11 Jul 2015

Keywords

  • Bovine tuberculosis
  • CD4 T cells
  • Immunology
  • Issue 101
  • Vaccine

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